Based on the requirements of Good Preparation Practice in Medical Institutions(GPP) and considering the actual conditions of the hospital,we decided upon a right way in the reconstruction of the hospital's preparation room and amelioration of the equipment and apparatus that constituted 30% of the investment.The results were reduced investment,higher efficiency,with perfect conformity to the GPP requirements.

Objective: To study the effect and mechanism of Rhodobryum giganteum (Schwaegr.) Par. 's anti-atherosclerotic effect. Methods:Vascular endothelial ceils were cultivated and the H2O2 were used to induce the oxidative stress injury of human umbilical vascular endothelial cells (HUVEC). Water extract of Rhodobryum giganteum (Schwaegr.) Par. Was added to cultivated HUVEC, and the activity of the cells was carefully determined by OD value with MTF method. The Griess Reagent was used to detect the NO concentration of different groups. At the same time, the NO fluorescence analysis probe, DAF-FM DA. (3-amino, 4-aminomethyl-2', 7'-difluorescein, diacetate), was used to determine the activity of NO synthase. Results:The most suitable stimulating concentration of H2O2 on HUVEC is 12.5 mmol· L-1. Water extract of Rhodobryum giganteum (Schwaegr.) Par. Was co-cultured with HUVEC damaged byH2O2 .The OD values indicate that 3.33,2.50 mg· mL-1 water extract groups increase activity of the cells significantly compared with the model group (P <0.05) ,and 3.33 mg·mL-1 is much better than 2.50 mg·mL-1 dose group. To determine the content of NO and NOS, the 2.50 mg·mL-1 dose has significant effect (P < 0.05). Conclusions: The H2O2 concentration of 12.5 mmol· L-1 could be used to establish the injured model of endothelial cell successfully. 2.50 and 3.33 mg·mL-1 water extract of Rhodobryum giganteum (Schwaegr.) Par. Have protective effect on ECV304 injured by H2O2. 2.50 mg· mL-1 water extract of Rhodobryurn giganteum (Schw aegr.) Par could increase the activity of NOS and promote the synthesis and secretion of NO.

Objectives: Observe the effects of acupuncture on obese SD rats fed by self-prescribed high calorie diet. 　Methods: The model group were male SD rats fed by self-prescribed high calorie diet, while the control group were male SD rats fed by common roughage. After acupuncture, the changes of some indexes were observed such as eating and drinking amount,body weight,length, Lee index,fattiness in groin and circumrenal areas, TG,TC,BG levels in blood etc.　 Results: The obesity was defined as the average body weight of the model group was 20% heavier than the average body weight in the control group. And the obesity ratio was 56%. The indexes were changed in different degrees with statistic significance after one period of treatment.　 Conclusions: Over-nutrition is one of the causes of obesity. Our high calorie feed for obese rat is reasonable. Acupuncture has obvious therapeutic effects for simple obesity.

BACKGROUND:Our previous study has already manifested that Chinese medicine Xiaokening can effectively prevent and treat the early diabetic nephropathy. OBJECTIVE:To compare the effect of rhein and archen on the apoptosis of mesangial cells of rats cultured with high glucose. METHODS:Mesangial cells were stimulated by different concentrations of rhein and archen (20, 40 and 80 μmol/L). Morphology of apoptotic cells was observed by hematoxylin-eosin staining. Karyon apoptosis was examined by fluorescence staining of DAPI. cellapoptosis rate was detected by flow cytometry. RESULTS AND CONCLUSION:The outcome of hematoxylin-eosin staining and DAPI staining indicated that the effect of rhein was much stronger than that of archen on the apoptosis of mesangial cells in the rats cultured with high glucose. Comparison of the apoptosis rate also indicated that the rhein had a stronger effect on the earlier and the later stage apoptotic rate of mesangial cells than archen. Both rhein and archen with different concentrations can induce apoptosis of mesangial cells, but the effect of rhein is much stronger.

BACKGROUND: Chinese medicines have better effects on preventing and treating diabetic nephropathy at early period, and the effects are caused by the diversity of the effective components of Chinese medicines which acts on the different targets of diabetic nephropathy.OBJECTIVE: To observe the effect of Xiaokening on the proliferation of mesangial cells under high glucose, and investigate the possible mechanism. DESIGN: A controlled observation.SETTING: Department of Endocrinology, the First Affiliated Hospital of Nanjing Medical University.MATERIALS: The experiments were completed in the Research Room of Cell Culture, Research Center of Endocrine Metabolism, the First Affiliated Hospital of Nanjing Medical University from July 2003 to May 2004. The rat mesangial cell lines HBZY-1 were bought from Chinese Center for Typical Culture Collection.METHODS: The mesangial cells were passaged and cultured according to the instructions. ① Grouping according to different concentration of stimulation: In the high glucose+Xiaokening group, Xiaokening of 6 concentrations were used, I.e., 20.0, 40.0, 60.0, 80.0, 120.0, 200.0 g/L. Meanwhile,normal glucose control group and high glucose control group were also set,the glucose concentration in the medium was 5.6 and 30 mmol/L respectively. The proliferations were observed after 72 hours. Grouping according to different time points of stimulation: There were normal glucose control group, high glucose control group and high glucose+Xiaokening group, the glucose concentration in the medium was 5.6, 30 and 30 mmol/L respectively, and the Xiaokening concentration was 60 g/L. The proliferations were observed at 24, 48 and 72 hours respectively in the three groups.② The dispensed cell suspension was placed into the 96-well plate, and 200 μL suspension for each well. The culture plate was placed in the inoculation box containing CO2 (0.05 in volume fraction) at 37 ℃ for 24 hours, then the supernatant was deserted, cell solutions containing different drugs of different concentrations were added in each group, 200 μL for each well. The 96-well plate was then placed in the culture box containing CO2 (0.05 in volume fraction) and 100% humidity at 37 ℃ for inoculation.In the groups of different concentrations, 20 μL MTT (5 g/L) was added into the wells after 72 hours. In the groups of different time points, 20 μL MTT (5 g/L) was added into the wells at 24, 48 and 72 hours. Then the plates were inoculated at 37 ℃ for 4 hours. Under microscope, it was observed that MTT get into the cells, the supernatant was sucked away, then 150 μL dimathyl sulfoxide was added to dissolve methyl alcohol, and shaken to even with plate rocking bed for 30 s, and the absorbance (A) values were recorded with the microplate reader at the wavelength of 492 nm.MAIN OUTCOME MEASURES: The proliferations of mesangial cells (A value) were observed at different time points and in Xiaokening of different concentrations.RESULTS: ①Proliferation of mesangial cells at different time points: The A values in the high glucose control group at 24, 48 and 72 hours (0.685 ±0.010, 0.750±0.087, 0.659±0.018) were higher than those in the normal glucose control group (0.586±0.054, 0.598±0.040, 0.527±0.047, P ＜ 0.05). In both the high and normal glucose control groups, the A value at 48 hours was higher than that at 24 hours (P ＜ 0.05), but the A value at 72 hours was lower than that at 48 hours (P ＜ 0.05). The A value in the high glucose+Xiaokening group at 72 hours was 0.538±0.023, and it was lower than that in the high glucose control group (P ＜ 0.05). ② Proliferation of mesangial cells in Xiaokening of different concentrations: Xiaokening high er than 60.0 g/L could obviously restrain the excessive stimulation of high glucose to the mesangial cells, and the effect was in a concentration-de pendent manner, but too high concentration (120.0 g/L) would result in the abscission and death of cells, whereas no survived cells could be observed in extremely high concentration (200.0 g/L). CONCLUSION: Xiaokening could inhibit the proliferation of mesan gial cells stimulated by high glucose after 72 hours in a concentration dependent manner, but too high concentration would cause strong cytotoxicity.

Objective:The best operation condition of extraction was found through analyzing the influences of both process of microwave pretreatment and hot wave washing. Methods: A new extraction method of chlorogenic acid was that the dry FLos Lonicerae was humidified with 75% ethanol-water solution at first, then the wet Flos Lonicerae was rapidly vaporized by microwave heating, at last chlorogenic acid was extracted after hot water washed twice, 10 minutes each time. Results: Compared with traditional extraction, the extraction time was about less than 6 times and then extraction ratio was increased by 1%. Conclusion: The method has high rate of extraction and short time of extraction.

Objective To establish an HPLC fingerprint of Herba Rhodobryi Rosei.Methods The fingerprint chromatography has been determined by RP-HPLC.The analysis was carried out with Dikma ODS C18 column(250 mm?4.6 mm,5 ?m).The mobile phase were:0.5% HAcCN(A),0.5% HAcH2O(B);Elution method:0-50 min,A was 20%-55%;50-60 min,A was 55%-95%;60-85 min,A was 95%-100%;keeping 5 min.Flow-rate was 1.0 mL/min.Wavelength was 360 nm and temperature was 30 ℃.It was analysized with the Estimating System of Similarity of 2004A Version(the Country's Pharmacopeia Committee)on the Chinese Medicine Fingerprint Chromatography.Results The fingerprint chromatography of Herba Rhodobryi Rosei was estabilished.Conclusion The method can be used in quality control of Herba Rhodobryi Rosei with accuracy and better repeatability.

Objective:To study the relationship between hepatic tissue insulin receptor substrate 2(IRS2) gene and type 2 diabetes,and the regulation of acupuncture on IRS2 gene expression of hepatic tissue in type 2 diabetes mellitus(T2DM) rats.Methods:Obese rats due to diet were injected in abdomen with small dose streptozotocin(STZ) to induce T2DM.Then these T2DM rats were stochastically divided into acupuncture group,Glibenclamide group and model group.After 4 weeks treatment,fasting blood sugar(FBS) was examined with active blood sugar meter,fasting insulin(FINS) was examined by radioimmunoassav,hepatic tissue IRS2mRNA was evaluated in real-time RT PCR,and normal rats were in control.Results:Serum levels of FBS(15.79?1.87) and FINS(37.20?3.92) in model group were higher than those in normal group(5.16?0.13,23.57?1.63),serum levels of FBS(5.46?0.20) and FINS(20.87?1.80) in acupuncture group were obviously lower than those in model group,which were similar to normal group.FBS(5.26?0.13) reduced obviously and serum FINS(28.10?1.86) reduced in some degree in Glibenclamide group compared with model.Relative contents of IRS2mRNA in model group were 2.19 folds compared with that in normal group.And in Glyburide group it was 4.59 folds compared with that in normal group.In acupuncture group it was 4.04% of that in the normal group.Conclusion:Abnormal hepatic tissue IRS2 mRNA expression may be one of pathogenesis of T2DM.Acupuncture can obviously depress hepatic tissue IRS2 mRNA expression,but Glibenclamide can not.

Objective To study the relationships among leptin, leptin receptor (OB-R) gene in arcuate nucleus and type 2 diabetes mellitus, and the effect of acupuncture on them. Methods Obese rats due to diet were injected in abdomen with small dosage streptozotocin to induce type 2 diabetes mellitus model, and were divided into acupuncture, glibenclamide and model group. Normal rats were in control. After treatment for 4 weeks, fasting blood sugar (FBS), fasting insulin (FINS), leptin, OB-R mRNA expression in arcuate nucleus were evaluated. Height, weight were measured and Lee's index were calculated before and after treatment. Results The FBS and FINS were higher in model group ［(15.79±1.87) mmol/L, (37.20±3.92) mIU/L］ than in control ［(5.16±0.13) mmol/L, (23.57±1.63) mIU/L］, but decreased in acupuncture group ［(5.46±0.20) mmol/L, (20.87±1.80) mIU/L］ and glibenclamide group ［(5.26±0.13) mmol/L, (28.10±1.86) mIU/L］. Leptin was lower in model ［(0.95±0.15) ng/ml］ than in control ［(1.32±0.15) ng/ml］, but was higher in glibenclamide group ［(1.83±0.23) ng/ml］ than in control. Weight and Lee's index decreased in model, acupuncture and glibenclamide groups after treatment. OB-R mRNA expression in arcuate nucleus in model wasn't different from control, acupuncture or glibenclamide group, nor between acupuncture and control, but was lower in glibenclamide group than in control. Conclusion Leptin decreased in type 2 diabetes mellitus rats when they became thin. Leptin resistance appears when treated with glibenclamide. Acupuncture may improve the OB-R mRNA expression in arcuate nucleus to some extent.

Purpose To investigate the expression and significance of HLA-G in ovarian serous carcinoma ( OSC) . Methods HLA-G antigen was immunohistochemically labeled on paraffin-embedded sections of 108 OSCs. The relationship between HLA-G expression and the clinicopathologic parameters was studied. Results The positive expression of HLA-G was observed in 58. 33% (63/108) of OSC tissues. Positive expression of HLA-G was significantly related with lymph node metastasis, recurrence, occurrence site, FIGO stage and MDACC grading system (P<0. 05). In survival analysis, the expression of HLA-G was significantly relevant to prognosis (P=0. 015). Multivariate Cox analysis showed the expression of HLA-G was an important prognosis factor of OSC (P=0. 01). Con-clusion The positive expression of HLA-G could predict high grade and advanced stage of OSC as well as poor prognosis. Also it could distinguish high-grade OSC from low-grade OSC.